Nitric Oxide Colorimetric Detection Kit

Sample Nitrite Conc. (uM) %CV Total NO Conc. (uM) %CV 1 45.1 4.4 70.5 6.8 2 73.3 9.1 107.4 4.4 3 132.7 1.3 157.8 1.8 Sample Nitrite Conc. (uM) %CV Total NO Conc. (uM) %CV Sample Nitrite Conc. (uM) %CV Total NO Conc. (uM) %CV 1 45.1 4.4 70.5 6.8 1 45.1 4.4 70.5 6.8 2 73.3 9.1 107.4 4.4 2 73.3 9.1 107.4 4.4 3 132.7 1.3 157.8 1.8 3 132.7 1.3 157.8 1.8 Sample Nitrite Conc. (uM) %CV Total NO Conc. (uM) %CV 1 44.1 3.1 68.8 7.4 2 66.4 4.0 112.1 5.7 3 126.7 6.3 154.4 4.1 Sample Nitrite Conc. (uM) %CV Total NO Conc. (uM) %CV Sample Nitrite Conc. (uM) %CV Total NO Conc. (uM) %CV 1 44.1 3.1 68.8 7.4 1 44.1 3.1 68.8 7.4 2 66.4 4.0 112.1 5.7 2 66.4 4.0 112.1 5.7 3 126.7 6.3 154.4 4.1 3 126.7 6.3 154.4 4.1

The DetectX® Nitric Oxide Detection Kit is designed to quantitatively measure Nitrate and Nitrite present in a variety of samples. Nitric Oxide content is derived from the sum of Nitrate (- NO3) and Nitrite (- NO2) . Please read the complete kit insert before performing this assay. Both Nitrate and Nitrite standards are provided to generate standard curves for the assay and all samples should be read off the appropriate standard curve. For Nitrite detection, samples are mixed with Color Reagents A and B and incubated at room temperature for 5 minutes. The colored product is read at 540 - 570 nm. The concentration of Nitrite in the sample is calculated, after making a suitable correction for any dilution of the sample, using software available with most plate readers. Total Nitric Oxide content is measured after the sample is incubated with Nitrate Reductase and NADH. The reductase in combination with NADH reduces Nitrate to Nitrite. After a 20 minute incubation at room temperature, Color Reagents A and B are added and incubated at room temperature for 5 minutes. The colored product is read and calculated as with the Nitrite determination above. The concentration of Nitrate in the sample is calculated by subtracting the measured Nitrite concentration from the Total Nitric Oxide concentration for the sample. This kit uses Nitrate and Nitrite Standard solutions calibrated to the US National Institute for Science and Technology Standard Reference Materials and ISO/IEC standards.

Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Reconstitution & Storage Instructions Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Antibody Array (AA) Protocol

Component Quantity Clear Half Area 96 well Plates 2 Plates Nitrate Standard 200 uL Nitrite Standard 200 uL Assay Buffer 60 mL NADH Concentrate 1.2 mL Nitrate Reductase 1 Vial Enzyme Stabilization Buffer 1 mL Color Reagent A 5 mL Color Reagent B 5 mL Component Quantity Component Quantity Clear Half Area 96 well Plates 2 Plates Clear Half Area 96 well Plates 2 Plates Nitrate Standard 200 uL Nitrate Standard 200 uL Nitrite Standard 200 uL Nitrite Standard 200 uL Assay Buffer 60 mL Assay Buffer 60 mL NADH Concentrate 1.2 mL NADH Concentrate 1.2 mL Nitrate Reductase 1 Vial Nitrate Reductase 1 Vial Enzyme Stabilization Buffer 1 mL Enzyme Stabilization Buffer 1 mL Color Reagent A 5 mL Color Reagent A 5 mL Color Reagent B 5 mL Color Reagent B 5 mL